Cited by 240Barrier Structure and Function StudiesCholesterol and Lipid MetabolismDrug Transport and Resistance Mechanisms

The DOI matches its Crossref record; 34 of the 34 references it cites were found in a registry or an index; 1 of the 28 sources its claims cite were checked for support.

Upregulation of the low density lipoprotein receptor at the blood-brain barrier: intercommunications between brain capillary endothelial cells and astrocytes.

Open access
Dehouck, B, Dehouck, M P, Fruchart, J C, Cecchelli, R·1994·The Journal of cell biology

Abstract

In contrast to the endothelial cells in large vessels where LDL receptors are downregulated, brain capillary endothelial cells in vivo express an LDL receptor. Using a cell culture model of the blood-brain barrier consisting of a coculture of brain capillary endothelial cells and astrocytes, we observed that the capacity of endothelial cells to bind LDL is enhanced threefold when cocultured with astrocytes. We next investigated the ability of astrocytes to modulate endothelial cell LDL receptor expression. We have shown that the lipid requirement of astrocytes increases the expression of endothelial cell LDL receptors. Experiments with dialysis membranes of different pore size showed that this effect is mediated by a soluble factor(s) with relative molecular mass somewhere between 3,500 and 14,000. Substituting astrocytes with smooth muscle cells or brain endothelium with endothelium from the aorta or the adrenal cortex did not enhance the luminal LDL receptor expression on endothelial cells, demonstrating the specificity of the interactions. This factor(s) is exclusively secreted by astrocytes cocultured with brain capillary endothelial cells, but it also upregulates the LDL receptor on other cell types. This study confirms the notion that the final fine tuning of cell differentiation is under local control.

Overview

AI snapshot

Written from the paper’s full text.

TL;DR

Brain capillary endothelial cells express LDL receptors at confluence despite high lipoprotein levels, unlike aortic endothelial cells, and coculture with astrocytes increases their LDL binding capacity. Astrocytes depleted of cholesterol release a soluble factor(s) of 3,500–14,000 mol wt that upregulates endothelial LDL receptor expression via new receptor synthesis.

Key findings

  • LDL binding to brain capillary endothelial cells was enhanced threefold when cocultured with astrocytes; maximal binding was 358 ng LDL/mg protein with astrocytes versus 215 ng LDL/mg protein without, with apparent Kd values of 43 nM versus 90 nM, respectively.
  • Cholesterol depletion of astrocytes by 36 h in lipoprotein-deficient serum decreased astrocyte cholesterol content by 32% (27.00 ± 3.37 vs 39.25 ± 4.03 µg cholesterol/mg protein, n = 6) and led to upregulation of endothelial LDL receptors, reaching a maximum of 600% of control binding at 4–5 h.
  • Astrocyte-conditioned medium increased LDL binding by 300% at 4–5 h; upregulation occurred with a 14,000 mol wt cutoff dialysis membrane but not with a 3,500 cutoff, indicating a soluble factor(s) between 3,500 and 14,000 mol wt.
  • The effect was specific: substituting astrocytes with smooth muscle cells or brain endothelium with aortic or adrenal cortex endothelium did not enhance luminal LDL receptor expression; acetylated LDL/scavenger receptor binding was not upregulated, and cycloheximide blocked the increase, indicating new LDL receptor synthesis.

Methods

  • Bovine brain capillary endothelial cells were cocultured with rat astrocytes on opposite sides of collagen-coated filter inserts; astrocytes were >95% GFAP positive. Bovine aortic endothelial cells, adrenal cortex endothelial cells, and smooth muscle cells were used for specificity comparisons.
  • LDL binding was performed at 4°C with [125I]LDL, with nonspecific binding determined by 20-fold excess unlabeled LDL; Scatchard analysis, C7 monoclonal antibody blocking, acetylated LDL binding, and HPLC cholesterol measurement were used.
  • Regulation experiments followed four phases: 12 d coculture in 15% calf serum, 36 h astrocyte incubation in 10% lipoprotein-deficient serum, induction with cholesterol-depleted astrocytes or conditioned medium at 37°C, then LDL binding at 4°C; dialysis membranes with 3,500 or 14,000 mol wt cutoffs and cycloheximide (10 µg/ml) were used in modified induction experiments.

Limitations

  • The paper states that the chemical nature of the astrocyte-regulating LDL receptor factor(s) is under investigation, that additional studies are underway to determine whether bound lipoproteins are degraded or transcytosed, and that 36 h LPDS incubation was used because cell lysis could be detected after 48 h.
  • The paper also notes that upregulation was lower with astrocyte-conditioned medium, possibly because the secreted molecule(s) is unstable.
  • Evident from the text: the findings rely on an in vitro bovine/rat coculture model, the soluble factor is not purified or chemically identified, and in vivo confirmation is not provided.

Model-written summary. Check it against the paper before relying on it.

Integrity signals

Registry

Crossref · 10.1083/jcb.126.2.465

Cites

34 of 34 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Verification

Claims the paper makes

1 of 28 cited sources checked · 0 supported · 0 flagged · accuracy not yet measured

Show all 59 claims

  • Probably not a substantial claim. 14
  • It may state two separate claims as one. 15
26 of 26
  • The paper relies on this without support.

    Aortic endothelial cells and smooth muscle cells have no intracellular pool of LDL receptor.

    “aortic ECs and smooth muscle cells have no intracellular pool of LDL receptor”
  • The paper relies on this without support.

    Astrocytes are capable of influencing the differentiation of brain endothelial cells in culture, as they are in vivo.

    “It is well known that astrocytes are capable of influencingthe differentiationofbrain ECs in culture, as they are in vivo.”
  • The paper relies on this without support.

    The direct action of LPDS on endothelial cells is well known, and lipoprotein receptor activity of endothelial cells is upregulated by exposure to LPDS after a minimum of 24 h.

    “the direct action of LPDS on endothelial cells is very well known, and lipoprotein receptor activity of EC is up- regulated by exposure to LPDS after a minimum of 24 h.”
  • The paper relies on this without support.

    Cycloheximide inhibits peptidyl transferase on the eukaryotic large ribosomal subunit and therefore protein synthesis in the cell.

    “Cycloheximide, known to inhibit peptidyl transferase on eu- karyiotic large ribosomal subunit, and therefore protein syn- thesis in the cell”
4 of 4

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Metadata

Authors
Dehouck, B., Dehouck, M. P., Fruchart, J. C., & Cecchelli, R.
Affiliations
Inserm (FR)
Venue
The Journal of cell biology
Published
1994-07-15
Volume
126
Issue
2
Pages
465-473
ISSN
  • 0021-9525 (ISSN-L)
  • 1540-8140
OpenAlex
W2159464926
Cited by
240
References
34
Citations by year
2026: 3 · 2025: 4 · 2024: 3 · 2023: 9 · 2022: 7 · 2021: 6 · 2020: 8 · 2019: 3 · 2018: 5 · 2017: 10 · 2016: 7 · 2015: 7 · 2014: 11 · 2013: 13 · 2012: 7
FWCI
7.04
Type
journal article
Language
en
Open access
Licence
cc-by-nc-sa
Locations
  • The Journal of Cell Biology (publishedVersion)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by-nc-sa)
  • PubMed Central (publishedVersion, cc-by-nc-sa)
  • PubMed (publishedVersion)
Topics
  • Life Sciences › Neuroscience › Neurology › Barrier Structure and Function Studies
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Biochemistry › Cholesterol and Lipid Metabolism
  • Health Sciences › Medicine › Oncology › Drug Transport and Resistance Mechanisms
Keywords
LDL receptor, blood-brain barrier, brain microvascular endothelial cells, astrocytes, LDL binding, soluble factors, receptor up-regulation, cell differentiation
MeSH terms
Adrenal Cortex, Animals, Aorta, Astrocytes, Binding Sites, Biological Factors, Blood-Brain Barrier, Brain, Capillaries, Cattle, Cell Communication, Cholesterol, Endothelium, Vascular, Kinetics, Lipoproteins, LDL, Muscle, Smooth, Receptors, LDL, Up-Regulation, Rats

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